Method Optimization and Detection of Aflatoxin Contaminants in Animal Feed Using an HPLC Fluorescence Detector Y Anastasia, R Widiastuti, R Maryam, Dalilah
Reasearch Center for Veterinary Science
Abstract
Aflatoxin contamination in animal feed is a major regulatory concern. Since feed is a complex analytical matrix, there is a need for validated methodologies for aflatoxins in feed. This study aimed to optimize a rapid and simple extraction technique to detect aflatoxins B1, B2, G1, and G2 in animal feed. The sample was extracted with acetonitrile and water (84:16, v/v), followed by cleaned up with solid phase extraction cartridges. The derived sample was then injected into the HPLC fluorescence detector, set at 355 nm and 368 nm wavelengths, also equipped with a C18 HPLC column at a flow rate of 1.0 mL/min. The results demonstrated an accuracy of 56% to 91.95% for each aflatoxin within the concentration range of 10-20 ng/mL, with precision below 5% relative standard deviation. Additionally, the linearity of concentrations ranging from 3.125-20 ng/mL exhibited a range of 0.9977-0.9994. The limit of detection ranged from 0.44-0.63 ng/mL, and the limit of quantification was 0.55-0.74 ng/mL. The optimized method was utilized to analyze 16 feed samples and 2 proficiency test samples, indicating its suitability for regulatory testing due to its simplicity, minimal equipment requirements, and cost-effectiveness, making it valuable for aflatoxin detection in animal feed.
